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. 2010 Jul 27;5(7):e11772. doi: 10.1371/journal.pone.0011772

Figure 1. bcl-2 modulation regulates HIF-1α protein expression in conditions strictly dependent on oxygen avaibility.

Figure 1

(A) Western blot analysis of HIF-1α and bcl-2 protein expression in total extracts of M14 cells transfected with siRNA targeting bcl-2 mRNA (si-bcl-2) or with a control scrambled si-RNA (si-contr) and then exposed to normoxia or hypoxia for 24 h. (B) Western blot analysis of HIF-1α and HIF-1β protein expression in total extracts of M14 control (puro) and bcl-2 stably overexpressing (Bcl2/5, Bcl2/37) cells plated under low (sparse) or high (dense) cell density conditions, or cultured under normoxia for 4 days or under hypoxia for 24 h. Western blot analysis of HIF-1α and HIF-1β protein expression in total extracts of the cells plated under high cell density conditions and (C) exposed to 24 h shaking or (D) cultured with different volumes of medium. (E) Western blot analysis of HIF-1α and HIF-1β protein expression in total extracts of cells exposed to Insulin (100 nM) or Epidermal Growth Factor (EGF, 20 ng/ml) for 24 h. (A–E) β-actin protein amounts are used to check equal loading and transfer of proteins. Western blot analyses representative of two independent experiments with similar results are shown.